title: Efficient siRNA transfection to the inner ear through the intact round window by a novel proteidic delivery technology in the chinchilla.
abstract: The use of small-interfering RNA (siRNA) has great potential for the development of drugs designed to knock down the expression of damage- or disease-causing genes. However, because of the high molecular weight and negative charge of siRNA, it is restricted from crossing the blood-cochlear barrier, which limits the concentration and size of molecules that are able to gain access to cells of the inner ear. Intratympanic approaches, which deliver siRNA to the middle ear, rely on permeation through the round window for access to the structures of the inner ear. We developed an innovative siRNA delivery recombination protein, TAT double-stranded RNA-binding domains (TAT-DRBDs), which can transfect Cy3-labeled siRNA into cells of the inner ear, including the inner and outer hair cells, crista ampullaris, macula utriculi and macula sacculi, through intact round-window permeation in the chinchilla in vivo, and there were no apparent morphological damages for the time of observation. We also found that Cy3-labeled siRNA could directly enter spiral ganglion neurons and the epithelium of the stria vascularis independently; however, the mechanism is unknown. Therefore, as a non-viral vector, TAT-DRBD is a good candidate for the delivery of double-stranded siRNAs for treating various inner ear ailments and preservation of hearing function.
authors.affiliation: Department of Otolaryngology, Huashan Hospital Fudan University, Shanghai, China.
authors.forename: W
authors.initials: W
authors.lastname: Qi
authors.affiliation: Center for Hearing and Deafness, State University of New York at Buffalo, Buffalo, NY, USA.
authors.forename: D
authors.initials: D
authors.lastname: Ding
authors.affiliation: Novoprotein Scientific Inc, Shanghai, China.
authors.forename: H
authors.initials: H
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: D
authors.initials: D
authors.lastname: Lu
authors.affiliation: Novoprotein Scientific Inc, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Novoprotein Scientific Inc, Shanghai, China.
authors.forename: J
authors.initials: J
authors.lastname: Ding
authors.affiliation: Department of Otolaryngology, Huashan Hospital Fudan University, Shanghai, China.
authors.forename: W
authors.initials: W
authors.lastname: Yan
authors.affiliation: Department of Otolaryngology, Huashan Hospital Fudan University, Shanghai, China.
authors.forename: M
authors.initials: M
authors.lastname: Jia
authors.affiliation: Department of Otolaryngology, Huashan Hospital Fudan University, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Guo
mesh_terms: D000818:Animals; D002232:Carbocyanines / Q000378:metabolism; D002682:Chinchilla; D005822:Genetic Vectors; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism*; D012405:Round Window, Ear / Q000378:metabolism*; D013136:Spiral Ganglion / Q000378:metabolism; D013316:Stria Vascularis / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D002232:Carbocyanines; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C089187:cyanine dye 3